Profiling histone post-translational modifications in mouse adult hepatocytes, biliary epithelial cells and biliary reprogrammed cells from hepatocytes.

Recent studies have demonstrated that hepatocytes can be reprogrammed into biliary epithelial cell-like cells under cholestatic liver injury. To investigate the changes in histone post-translational modification in hepatocyte-derived reprogramming cells, we performed Cleavage Under Targets and Tagmentation (CUT&Tag)-sequencing. Rosa26-LSL-Cas9-EGFP mice were injected with serotype 8 adeno-associated viruses (AAV8) packaged with TBG-Cre (AAV8-TBG-Cre) to genetically label adult hepatocytes. Mice were then treated with 0.1% 3,5-diethoxycarbonyl-1,4-dihydrocollidine (DDC)-containing diet to induce cholestatic liver injury for 8-9 weeks. Reprogrammed cells in early (EGFP+ CD24-), intermidate (EGFP+ CD24+ EPCAM-) and late stages (EGFP+ CD24+ EPCAM+) as well as and biliary cells (EGFP- EPCAM+) were harvested by FACS. Normal hepatocyts, as a control group, were harvested by the percoll density gradient method from mice maintained on normal diet.